ST 37 And ST 25 Induce Different Effects On Colonic Motility Via The Enteric Nervous System

Objective:Based on a novel mouse constipation model,the effect of electroacupuncture on the regulation of intestinal motility in ST 37 and ST 25 were investigated.The changes of nerve marker and different types of neurons in the enteric nervous system were observed.To identify regulation of colonic motility of the local nerve mechanism and for the clinical rational use of specific acupuncture treatment of intestinal disease to provide experimental basis.Methods:Part I:Methods of mouse constipation model.C57BL/6J mice were randomly divided into three groups:a 0-4? saline-treated group(group A),a normal feeding group(group B),and a 15-20? saline-treated group(group C),randomly numbered,and raised in single cages that allowed normal access to food and water.Wire netting was used to facilitate the separation and collection of stools.To eliminate the influence of the biological rhythms,intragastric administrations were conducted at 8:00 am once daily for 14 d.Mice in ice-cold and saline control groups were initially administered either ice-cold or room-temperature saline,respectively,at a dose of 0.2 mL/mouse,and then the dose increased by 0.05 mL/mouse every 5 d.Blank control group mice were raised normally without intragastric administration.Animals were normally raised and monitored for an additional 6 d after the cessation of the intragastric administrations.Part II:Constipation mice were randomly divided into three groups:model group,ST 37 group and ST 25 group.The same number of normal mice were set as control group.The parameters were 2/15 Hz,1 mA,and the time were 15 min/d for 3 days.In the recording of intestinal electrophysiological experiments,constipation mice and normal mice were selected.The self-made water bladder was placed in the proximal colon or distal colorectal,after the emergence of the waveform to be stable,first save a section for the baseline reference value,and then random order of electro-acupuncture stimulation,and the the next point.The parameters were 2/15 Hz,1 mA,and the time was 1 min._Through different experiments,the effects of ST 37 and ST 25 on constipation model were analyzed.Part ?:Constipation mice were randomly divided into three groups:model group,ST 37 group and ST 25 group.The same number of normal mice were set as control group.After the intervention of 3d,fasting for 12h to remove the proximal colon tissue.The expressions of PGP 9.5 and cholinergic neuron markers ChAT and nitrogen neuron marker nNOS were detected by immunohistochemistry,qPCR and Western Blot.Analysis of ST37 and ST25 produced differences in the effect of the intestinal nerve mechanism.Results:Part ?:Based on the percentage of stool weight changed compared with baseline(before irritation)in 9-14 d,stool weight changed were classified into three levels.Each level showed a different body state,in state ?,the stools became small and dry..This constipation model(State ?)only required a short intervention cycle(14 d)and normalcy was achieved 6 d after the termination of irritation,which is convenient and practical.The change of stool weight and size may serve as a useful tool to judge a constipation model success or not.Part ?:ST 37 and ST 25 were both effective,but the effects were not completely consistent.In the first discharge indicator experiment,ST 37 recovered to normal level,while ST 25 failed to return to normal.In terms of stool weight and form,ST 25 group was still lower than that in normal and ST 37 group.In addition,in the intestine pressure experiment,ST 37 and ST 25 can improve the colorectal pressure,and ST 37 could improve the proximal colonic pressure,while ST 25 had no effect.Part ?:The results of immunohistochemistry showed that ST 37 could regulate the expression of ENS in the proximal colon of constipated mice,but ST 25 had no effect.In addition,qPCR and Western Blot showed that ST 37 could down-regulate the expression of nNOS to the normal level,and up-regulate the expression of ChAT to normal level,while ST 25 could only be down-regulated the expression of nNOS to normal levels.Conclusion:1.Irritation with ice-cold saline is a stable,repeatable method in building constipation model in mice for exploring the pathogenesis and treatment options of constipation,2.ST 37 and ST 25 are both effective,but the effects are not the same.ST 37 and ST 25 could enhance the colorectal pressure,and ST 37 could enhance the proximal colonic pressure.3.ST 37 and ST 25 induce different effects on colonic motility via the enteric nervous system by affecting excitatory and inhibitory neurons.